Fig. 2. Indirect immunofluorescence analysis of HA-tagged lamin A (LaA, A-J) and lamin C (LaC, K-O) alleles expressed in HeLa cells. In all cases the cells were labeled with the 12CA5 anti-HA antibody. F-J also include DNA staining with Höchst dye 33258. In the case of the lamin A mutants only LaA N195K (C and H) exhibits a distribution that is overtly different to wild-type (LaA WT, A and F). This protein remains largely nucleoplasmic and gives rise to intranuclear inclusions or aggregates in 25% of transfected cells (insets). These most likely correspond to those cells with the highest LaA N195K expression levels. Of the lamin C alleles, only the wild-type protein, LaC WT (K) and LaC R482W (N) are appropriately targeted to the nuclear envelope. The remainder localize exclusively to the nucleoplasm (L, M and O).